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101.
Enhancement of hyperthermia-induced apoptosis by local anesthetics on human histiocytic lymphoma U937 cells 总被引:2,自引:0,他引:2
Arai Y Kondo T Tanabe K Zhao QL Li FJ Ogawa R Li M Kasuya M 《The Journal of biological chemistry》2002,277(21):18986-18993
The combined effects of hyperthermia at 44 degrees C and local anesthetics on apoptosis in human histiocytic lymphoma U937 cells were investigated. When the cells were exposed to hyperthermia for l0 min marginal DNA fragmentation and nuclear fragmentation were observed. In the presence of amide-type local anesthetics further enhancement was found depending on concentration. The order of the concentration required for maximum induction was the reverse order of the lipophilicity (prilocaine > lidocaine > bupivacaine). Western blotting revealed that in hyperthermia there was initial release of Ca(2+) from the intracellular store site as indicated by increased expression of the type 1 inositol-1,4,5-trisphosphate receptor. However, the combination with lidocaine did not induce any further enhancement. Lidocaine enhanced the decrease in ATP content and the increase in intracellular Ca(2+) concentration in individual cells induced by hyperthermia. In addition, superoxide formation, decrease in the mitochondrial membrane potential, and activation of intracellular caspase-3 were found in the cells treated with hyperthermia and lidocaine. All of these were suppressed in part in the presence of the intracellular Ca(2+) ion chelator BAPTA-AM (bis-(O-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl). The present results indicate that local anesthetics at optimal concentrations enhance hyperthermia-induced apoptosis via Ca(2+)- and mitochondria-dependent pathways. Initial release of Ca(2+) from intracellular store sites caused by hyperthermia and followed by the subsequent increase in the intracellular Ca(2+) concentration and the additional activation of the mitochondrial caspase-dependent pathway (partly regulated by intracellular Ca(2+) concentration) plays a crucial role in the enhancement of apoptosis induced by the combination of hyperthermia and lidocaine. 相似文献
102.
103.
Immobilization of oligodeoxyribonucleotides with multiple anchors to microchips 总被引:5,自引:3,他引:5 下载免费PDF全文
Facile modification of oligodeoxyribonucleotides is required for efficient immobilization to a pre-activated glass surface. This report presents an oligodeoxyribonucleotide which contains a hairpin stem–loop structure with multiple phosphorothioate moieties in the loop. These moieties are used to anchor the oligo to glass slides that are pre-activated with bromoacetamidopropylsilane. The efficiency of the attachment reaction was improved by increasing the number of phosphorothioates in the loop, as shown in the remarkable enhancement of template hybridization and single base extension through catalysis by DNA polymerase. The loop and stem presumably serve as lateral spacers between neighboring oligodeoxyribonucleotides and as a linker arm between the glass surface and the single-stranded sequence of interest. The oligodeoxyribonucleotides of this hairpin stem–loop architecture with multiple phosphorothioate moieties have broad application in DNA chip-based gene analysis. 相似文献
104.
KPC1 (Kip1 ubiquitylation-promoting complex 1) is the catalytic subunit of the ubiquitin ligase KPC, which regulates the degradation
of the cyclin-dependent kinase inhibitor p27kip1 at the G1 phase of the cell cycle. To elucidate the expression and role of KPC1 in nervous system lesion and repair, we performed
an acute spinal cord contusion injury (SCI) model in adult rats. Western blot analysis showed a significant up-regulation
of KPC1 and a concomitant down-regulation of p27kip1 following spinal injury. Immunohistochemistry and immunofluorescence revealed wide expression of KPC1 in the spinal cord,
including expression in neurons and astrocytes. After injury, KPC1 expression was increased predominantly in astrocytes, which
highly expressed PCNA, a marker for proliferating cells. Co-immunoprecipitation demonstrated increased interactions between
p27kip1 and KPC1 4 days after injury. To understand whether KPC1 plays a role in astrocyte proliferation, we applied LPS to induce
astrocyte proliferation in vitro. Western blot analysis demonstrated that p27kip1 expression was negatively correlated with KPC1 expression following LPS stimulation. Immunofluorescence analysis showed subcellular
localizations of p27kip1 and KPC1 were also changed following the stimulation of astrocytes with LPS. These results suggest that KPC1 is related to
the down-regulation of p27kip1; this event may be involved in the proliferation of astrocytes after SCI. 相似文献
105.
Determination of genetic relationships among five indigenous Chinese goat breeds with six microsatellite markers 总被引:16,自引:0,他引:16
Microsatellite variation was analyzed in five Chinese indigenous goat breeds, which include four Cashmere breeds (Tibetan, Neimonggol, Liaoning, Taihang) and one Hubei local breed (Matou) used for meat production. Five ovine and one bovine microsatellites, selected from eight ovine microsatellites and five bovine microsatellites were suitable for use in this study. With these six microsatellites, allele frequencies, heterozygosity, polymorphism information content (PIC) and effective allele number were calculated. A neighbor-joining tree was constructed using Nei's standard genetic distance (1978). In the tree, Neimonggol and Liaoning were grouped together, then with Taihang; while Tibetan and Matou individually had their own branch. The genetic relationship of five breeds corresponds to their history and geographic origins. 相似文献
106.
107.
The model conjugates phycocyanin-allophycocyanin (C-PC-APC) and phycoerythrocyanin-phycocyanin-allophycocyanin (PEC-C-PC-APC)
were synthesized by using a heterobifunctional coupling reagent N-succinimidyl-3-(2-pyridyldithio)propionate. The rod-core
complex (αβ)6
PCLRC
27(αβ)3
APCLC
8.9 and phycobilisomes were separated from Anabaena variabilis. Energy transfer features for the conjugates and the complexes
were compared. The absorption and fluorescence emission spectra indicated that the linker-peptides mediate interaction of
phycobiliproteins and prompt energy transfer. The energy transfer in the conjugates was detected by fluorescence emission
spectra and confirmed by the addition of dithiothreitol. The conjugates may be used as models for studying the energy transfer
mechanism in phycobilisomes.
This revised version was published online in September 2006 with corrections to the Cover Date. 相似文献
108.
It has been known that Rho-associated protein kinase (ROCK) signaling regulates the migration of vascular smooth muscle cells (VSMCs). However, the isoform-specific roles of ROCK and its underlying mechanism in VSMC migration are not well understood. The current study thus aimed to investigate the roles of ROCK1/2 and their relationship to the MAPK signaling pathway in platelet-derived growth factor (PDGF)-induced rat aorta VSMC migration by manipulating ROCK gene expression. The results revealed that ROCK1 small interfering ribonucleic acid (siRNA) rather than ROCK2 siRNA decreased PDGF-BB-generated VSMC migration, and upregulation of ROCK1 expression via transfection of constructed pEGFP-C1/ROCK1 plasmid further increased the migration of PDGF-BB-treated VSMCs. In PDGF-treated VSMCs, ROCK1 siRNA did not affect the phosphorylation levels of ERK and p38 in the cytoplasm, but decreased the level of ERK phosphorylation in the nucleus. These findings demonstrate that activated ROCK1 can promote VSMC migration through facilitating phosphorylation and nuclear translocation of ERK protein. 相似文献
109.
Marjan Huizing Rangaprasad Sarangarajan Erin Strovel Yang Zhao William A. Gahl Raymond
E. Boissy 《Molecular biology of the cell》2001,12(7):2075-2085
Patients with Hermansky-Pudlak syndrome type 2 (HPS-2) have mutations in the beta 3A subunit of adaptor complex-3 (AP-3) and functional deficiency of this complex. AP-3 serves as a coat protein in the formation of new vesicles, including, apparently, the platelet's dense body and the melanocyte's melanosome. We used HPS-2 melanocytes in culture to determine the role of AP-3 in the trafficking of the melanogenic proteins tyrosinase and tyrosinase-related protein-1 (TRP-1). TRP-1 displayed a typical melanosomal pattern in both normal and HPS-2 melanocytes. In contrast, tyrosinase exhibited a melanosomal (i.e., perinuclear and dendritic) pattern in normal cells but only a perinuclear pattern in the HPS-2 melanocytes. In addition, tyrosinase exhibited a normal pattern of expression in HPS-2 melanocytes transfected with a cDNA encoding the beta 3A subunit of the AP-3 complex. This suggests a role for AP-3 in the normal trafficking of tyrosinase to premelanosomes, consistent with the presence of a dileucine recognition signal in the C-terminal portion of the tyrosinase molecule. In the AP-3-deficient cells, tyrosinase was also present in structures resembling late endosomes or multivesicular bodies; these vesicles contained exvaginations devoid of tyrosinase. This suggests that, under normal circumstances, AP-3 may act on multivesicular bodies to form tyrosinase-containing vesicles destined to fuse with premelanosomes. Finally, our studies demonstrate that tyrosinase and TRP-1 use different mechanisms to reach their premelanosomal destination. 相似文献
110.
本文对项青霉D_(1(?))的四个木聚糖酶组分的特性进行了研究。木聚糖酶组分D_(x1)、D_(x4)的最佳反应pH为4.8,最适温度分别为40℃和50℃,D_(x2)和D_(x3)的最适pH和温度都分别为pH4.2和50℃。Ag~(++)、Hg~(++),Cu~(++)对四个组分的活性均有强烈的抑制作用,SDS也能产生明显的抑制效果。Mn~(++)对D_(x1)具有促进作用。D_(x1)、D_(x4)在以燕麦木聚糖为底物时活性最高,其Km值分别为11.7(mg/ml)和8.3(mg/ml),D_(x2)和D_(x3)则分别在水解红麻杆木聚糖和落叶松木聚糖时活性最强,Km值分别为8.4(mg/ml)和6.3(mg/ml)。水解燕麦木聚糖,D_(x1)的产物主要为木糖,同时带有少量的低聚木糖。D_(x2)、D_(x3)和D_(x4)的产物则包括木糖和较多的低聚木糖。D_(x4)与D_(x2)及D_(x3)之间在水解燕麦木聚糖时存在协同作用关系。 相似文献